Introducing the new anti-mCherry antibody

Because of its brightness, superior photostability, and rapid maturation rate, the mCherry monomeric red fluorescent protein is widely used for monitoring physiological processes and detecting transgenic expression. To expand the options for researchers using the mCherry red fluorescent protein, we now provide an anti-mCherry rat monoclonal antibody that recognizes both native and denatured forms of mCherry and mCherry fusion proteins; no cross-reactivity was demonstrated against several forms of GFP and RFP. This affinity-purified rat monoclonal IgG2a has been validated for use in western blot, immunocytochemistry (including imaging and flow cytometry), and immunoprecipitation applications.

FxCycle™ PI/RNase Staining Solution is an optimized mixture of propidium iodide and DNase-free RNase A for the flow cytometric analysis of DNA content in fixed cells. This ready-to-use solution is specially formulated with a permeabilization reagent in DPBS. Just add the staining solution to fixed cells, incubate at room temperature, and acquire data on a flow cytometer; no washing steps are required.

Propidium iodide (PI), a popular red-fluorescent nucleic acid stain, binds to DNA by intercalating between bases with little or no sequence preference and a stoichiometry of one dye per 4–5 base pairs. PI also binds to RNA, necessitating treatment with nucleases to distinguish between RNA and DNA staining.

Design and implement your own Qubit® assays with MyQubit

MyQubit firmware brings your favorite fluorescence assays right to your benchtop, providing a simple and reliable platform for diverse quantitation needs—from basic research and quality control to process monitoring and beyond. Any reagent or assay that is spectrally compatible with the Qubit® hardware can be adapted for use with the Qubit® 2.0 Fluorometer using the MyQubit firmware, which comes preloaded on all new instruments or can be downloaded from the Qubit® web page.

Because the Qubit® 2.0 Fluorometer is operated by simple components, the creation of new applications is as straightforward as matching the spectral characteristics of the assay with the instrument’s LEDs and emission filters. You can use existing Life Technologies kits and reagents as the basis for new Qubit® assays, or develop novel quantitative assays. To get you started, we have created three MyQubit assays using Molecular Probes® reagents:

MyQubit Amplex® Red cholesterol assay

MyQubit Amplex® Red glucose assay

MyQubit microRNA (miRNA) assay

You can upload one or more of these assay files to your Qubit® 2.0 Fluorometer from the Qubit® web page, or get started on your own Qubit® assay with the MyQubit firmware.

Comparison of detection techniques for quantitation of small RNA in the presence of ribosomal RNA. rRNA at the concentrations listed was spiked into solutions containing 2 ng/µL siRNA, then read using the MyQubit miRNA assay (equivalent to the Qubit® microRNA assay) or the Qubit® RNA assay, or by 260 nm absorbance (A260) on the NanoDrop® spectrophotometer.

The GeneArt® Gene Synthesis Kit with CorrectASE™ error correction technology provides all the reagents you need for successful do-it-yourself gene synthesis, typically in just 3 days from oligonucleotide assembly to sequence-verified clone. High-fidelity Platinum® Pfx DNA Polymerase with automatic hot start facilitates oligonucleotide assembly and gene or fragment amplification prior to and after error correction. Quant-iT™ PicoGreen® dsDNA Reagent, an ultrasensitive fluorescent nucleic acid stain, enables accurate quantitation of dsDNA, which is essential for achieving the ideal DNA:CorrectASE™ enzyme ratio. The resulting PCR fragments can be cloned into the pCR™-Blunt II-TOPO® vector in a simple 5-minute reaction; this gene synthesis system is compatible with many cloning technologies, including GeneArt® seamless cloning and Gateway® cloning.

The CorrectASE™ enzyme, also available as a stand-alone reagent, removes mismatches caused by oligonuceotide synthesis errors, leading to a 3- to 10-fold reduction in mutations in synthetic genes or fragments. Depending on the source, commercially available synthetic oligonuceotides can exhibit high error rates—ranging from one in every 1,000 bases to one in every 300 bases. These errors cause frameshift (deletion or insertion) and substitution mutations during gene synthesis. The CorrectASE™ enzyme is introduced into the incubation step after the initial PCR assembly of oligonucleotides. The PCR products are denatured and reannealed so that any mutations will be mismatched. The CorrectASE™ enzyme binds to these mismatches and nicks both DNA strands 3´ of the error, and then the 3´→ 5´ exonuclease activity of the enzyme removes the errors. A final PCR amplification with a proofreading polymerase assembles the corrected fragments, thus increasing the likelihood of isolating clones with the proper sequence. Typically, only 2–4 clones need to be screened, depending on the initial oligonucleotide quality, as compared with 10–16 clones in a workflow that does not include an error correction step. Thus, both labor time and sequencing costs can be decreased when you incorporate CorrectASE™ error correction technology into your gene synthesis workflow.